Project description:Uncovering the mechanisms-based quality control of traditional Chinese medicine against essential hypertension by network analysis and experimental validation
Project description:The present invention relates to methods for determining soil quality, and especially soil pollution, using the invertebrate soil organism Folsomia candida also designated as springtail. Specifically, the present invention relates to a method for determining soil quality comprising: contacting Folsomia Candida with a soil sample to be analysed during a time period of 1 to 5 days; isolating said soil contacted Folsomia Candida; extracting RNA from said isolated soil contacted Folsomia Candida; determing a gene expression profile based on said extracted RNA using microarray technology; comparing said gene expression profile with a reference gene expression profile; and determing soil quality based expression level differences between said gene expression profile and said control expression profile.
Project description:Quality control (QC) in mass spectrometry (MS)-based proteomics is mainly based on data-dependent acquisition (DDA) analysis of standard samples. Here, we collected 2638 files acquired by data independent acquisition (DIA) and paired DDA files from mouse liver digests using 21 mass spectrometers across nine laboratories over 31 months. Our data demonstrated that DIA-based LC-MS/MS-related consensus QC metrics exhibit higher sensitivity compared to DDA-based QC metrics in detecting changes in LC-MS status. We then optimized 15 metrics and invited 21 experts to manually assess the quality of 2638 DIA files based on those metrics. Based on the annotation results, we developed an AI model for DIA-based QC in the training set of 2110 DIA files. This model predicted the liquid chromatography (LC) performance with an AUC of 0.91 and the MS performance with an AUC of 0.97 in an independent validation dataset (n = 528). Finally, we developed an offline software called iDIA-QC for convenient adoption of this methodology for LC-MS QC.
Project description:Quality control (QC) in mass spectrometry (MS)-based proteomics is mainly based on data-dependent acquisition (DDA) analysis of standard samples. Here, we collected 2638 files acquired by data independent acquisition (DIA) and paired DDA files from mouse liver digests using 21 mass spectrometers across nine laboratories over 31 months. Our data demonstrated that DIA-based LC-MS/MS-related consensus QC metric exhibit higher sensitivity compared to DDA-based QC metric in detecting changes in LC-MS status. We then optimized 15 metrics and invited 21 experts to manually assess the quality of 2638 DIA files based on those metrics. Based on the annotation results, we developed an AI model for DIA-based QC in the training set of 2110 DIA files. This model predicted the liquid chromatography (LC) performance with an AUC of 0.91 and the MS performance with an AUC of 0.97 in an independent validation dataset (n = 528). Finally, we developed an offline software called iDIA-QC for convenient adoption of this methodology for LC-MS QC.
Project description:Quality control (QC) in mass spectrometry (MS)-based proteomics is mainly based on data-dependent acquisition (DDA) analysis of standard samples. Here, we collected 2638 files acquired by data independent acquisition (DIA) and paired DDA files from mouse liver digests using 21 mass spectrometers across nine laboratories over 31 months. Our data showed that DIA-based LC-MS/MS related consensus QC metric is more sensitive than DDA-based QC in detecting MS status changes. We then optimized 15 DIA-QC metrics, and invited to manually assess the quality of 2638 DIA files generated by 21 mass spectrometers based on each metric. Based on the annotation results, we developed an AI model for DIA-based QC in the training set of 2059 DIA files, and predicted the liquid chromatography (LC) performance with an AUC of 0.91 and the MS performance with an AUC of 0.97 in an independent validation dataset (n = 523). Finally, we developed an offline software called iDIA-QC for convenient adoption of this methodology for LC-MS QC
Project description:The European clam, Ruditapes decussatus (Linnaeus, 1758) is a bivalve mollusc of the family Veneridae native to the European Atlantic and Mediterranean coastal waters. Its production is exclusively based on natural recruitment, which is subject to high annual fluctuations due to adversely affected by pollution and other environmental factors. A microarray-based analysis was performed with the objectives of describe genomic feature of oocytes and identify potential markers of oocyte quality in the economically important European clam, Ruditapes decussatus. The oocytes of a total of 25 females from Ria de Aveiro, Western coast of Portugal, were selected for this study and their quality was estimated by early developmental success until D-larval rate, under controlled conditions.
Project description:Currently, batch release of toxoid vaccines, such as diphtheria and tetanus toxoid, requires animal tests to confirm safety and immunogenicity. Efforts are being made to replace these tests with in vitro assays in a consistency approach. Limitations of current in vitro assays include the need for reference antigens and most are only applicable to drug substance, not to the aluminum adjuvant-containing and often multivalent drug product. To overcome these issues, a new assay was developed based on mimicking the proteolytic degradation processes in antigen-presenting cells with recombinant cathepsin S, followed by absolute quantification of the formed peptides by liquid chromatography-mass spectrometry. Temperature-exposed tetanus toxoids from several manufacturers were used as aberrant samples and could easily be distinguished from the untreated controls by using the newly developed degradomics assay. Consistency of various batches of a single manufacturer could also be determined. Moreover, the assay was shown to be applicable to Al(OH)3 and AlPO4-adsorbed tetanus toxoids. Overall, the assay shows potential for use in both stability studies and as an alternative for in vivo potency studies by showing batch-to-batch consistency of bulk toxoids as well as for aluminum-containing vaccines.
Project description:Mass spectrometry (MS)-based single-cell proteomics (SCP) provides us the opportunity to unbiasedly explore biological variability within cells without the limitation of antibody availability. This field is rapidly developed with the main focuses on instrument advancement, sample preparation refinement, and signal boosting methods; however, the optimal data processing and analysis are rarely investigated which holds an arduous challenge because of the high proportion of missing values and batch effect. Here, we introduced a quantification quality control to intensify the identification of differentially expressed proteins (DEPs) by considering both within and across SCP data. Combining quantification quality control with isobaric match between runs (IMBR) and PSM-level normalization, an additional 12% and 19% of proteins and peptides, regarding to more than 90% of proteins/peptides contained valid value, were quantified. Clearly, quantification quality control was able to reduce quantification variations and q-values with the more apparent cell type separations. In addition, we found that PSM-level normalization performed similarly to other protein level normalization but kept the original data profiles without the additional requirement of data manipulation. In proof of concept of our refined pipeline, six uniquely identified DEPs exhibiting varied fold-changes and playing critical roles for melanoma and monocyte functionalities were selected for validation using immunoblotting. Five out of six validated DEPs showed an identical trend with the SCP dataset, emphasizing the feasibility of combining the IMBR, cell quality control, and PSM-level normalization in SCP analysis, which is beneficial for future SCP studies.